Characterisation of TK6 cell line during direct adaptation to chemically defined serum replacement (CDSR) supplemented media. Cell viability (A), cell count (C), and cell size (D) were measured every cell passage and pooled every three passages and expressed as mean (±SD). Mitochondrial activity (bars) and cytotoxicity (dots) (B) were measured at indicated on x-axis passage number and expressed as mean (±SD). Dotted lines show the highest and/ or lowest values of cell viability (A), growth (C), and cell diameter (D) of cells growing in 10% (v/v) FBS-supplemented medium. Significant difference between conditions was analysed by two-way ANOVA. Statistical significance was indicated as follows: for (B): cytotoxicity statistical significance was indicated with *: * indicates p < 0.05 and *** indicate p < 0.001; while mitochondrial activity statistical significance was indicated with #: ### indicate p < 0.05; for (C): * indicates p < 0.05 and ** indicate p < 0.01.

Contexts in source publication

Context 1

... cells were directly transferred from 10% v/v FBSsupplemented media to 10% v/v CDSR-supplemented media. Subsequently cell proliferation, cell diameter and viability (including mitochondrial activity and cytotoxicity), were monitored for 20 passages (10 weeks) (Figure 2). Viability and cell diameter of TK6 cells in 10% v/v CDSR-supplemented medium were not significantly different (p < 0.05) from cells cultured in 10% v/v FBS supplemented medium (Figures 2A, D). ...

Context 2

... cell proliferation, cell diameter and viability (including mitochondrial activity and cytotoxicity), were monitored for 20 passages (10 weeks) (Figure 2). Viability and cell diameter of TK6 cells in 10% v/v CDSR-supplemented medium were not significantly different (p < 0.05) from cells cultured in 10% v/v FBS supplemented medium (Figures 2A, D). However, the viability of TK6 cells directly transferred cells at each passage was continually below the acceptance threshold (Figure 2A). ...

Context 3

... and cell diameter of TK6 cells in 10% v/v CDSR-supplemented medium were not significantly different (p < 0.05) from cells cultured in 10% v/v FBS supplemented medium (Figures 2A, D). However, the viability of TK6 cells directly transferred cells at each passage was continually below the acceptance threshold (Figure 2A). Additionally, cytotoxicity of TK6 cells in 10% v/v CDSRsupplemented medium was significantly lower at passage 4 (p < 0.001), passage 9 (p < 0.05) and passage 13 (p < 0.001) when compared with standard FBS-containing culture, while significantly lower mitochondrial activity was only observed at passage 4 (p < Significance between conditions was assessed using a one-way ANOVA. ...

Context 4

... ( Figure 2B). Furthermore, a significant (p < 0.05) reduction in cell number (indicating reduced proliferation) was reported at each passage ( Figure 2C). ...

Context 5

... Figure 2B). Furthermore, a significant (p < 0.05) reduction in cell number (indicating reduced proliferation) was reported at each passage ( Figure 2C). Cell proliferation and viability of cells directly adapted to CDSR was below the acceptance threshold and hence this transition methodology was discontinued. ...

Context 6

... the optimisation of medium composition and adaptation protocol is specific for each cell line making this an onerous process. In this study, we explored two adaptation methodologies i) direct adaptation, where cells were transferred from 10% (v/v) FBS containing medium directly to 0% (v/v) containing medium during one sub-culture (Figure 2) and ii) gradual adaptation, where cells where gradually sub-cultured using culture medium containing reduced concentrations until FBS was completely substituted for CDRS 0% (v/v) (Figure 3). Whilst both methodologies maintained TK6 cell growth in animal-free CDSR medium, the proliferation rate of TK6 cells was significantly reduced using the direct adaptation methodology and hence the gradual adaptation methodology was selected to investigate the long-term effects of CDSR-supplemented medium on TK6 cell properties. ...